Currently I'm trying to adjust Oil Red O staining method for liver lipids. Base protocol is A. Mehlem's (Nature Protocols; Vol. 8, No. 6; 2013; p. 1149-1154). I tried different times of ORO staining and counterstaining with hematoxylin. Had some problems with non-specific staining of cracks between cells which might have been solved with varying times. As I believe in the pics the darker dots are lipids and red colour between cells is other type of lipids? Maybe someone who has experience with ORO staining method could explain if attached pictures looks good and if not maybe suggest some corrections which might improve staining?
Liver samples are from chickens treated with high concentration of TBT.
Sample 7: 5 min. in ORO, 15 s. HTX, 30 min. wash; S8: 5 min. ORO, 1 m. HTX, 60 min. wash; S12: 5 min. ORO, 3 min. HTX, 30 min wash; S13: 5 min. ORO, 1 min. HTX, 30 min. wash.
Dear Andre,
I think the cracks, which you mention between the hepatocytes may be the sinusoids, partly filled with erythrocytes. I assume that your Oil red staining (large one) is O.K.
Yours sincerely
Daniela
This oil red o staining is fine. I want to ask you, are you using 10% formaline for fixing the frozen sections in the slides.
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